sk:i’’rimijb;it
A c Y CLI c
I!J(il [CUNTRIBUTION XROM
THE
s u c; .z 11 s u C L I:OSID 15 AS .z LOG s
DEPARTMENT O F CHEMISTRY
OF
3095
THEOHIO STATE UNIVERSITY]
Acyclic Sugar Nucleoside Analogs’ 11. L. WOLFROM, A. B. FOSTER, P. McWAIN, W. VON BEBENBURG,
A. THOMPSON
AND
Received Februanj 3, 1961 T h e preparation and characterization of a pair of l-epimers of 1-(9-aden~l)-l-deo?ry-l-O-methyl-aldeh~d~~-galactose aldehydrol ( I X ) and of 9-8-D-galactopyranosyhdenine are described.
The hydroxyl groups of the aldehydrol form of the acyclic aldose acetates readily undergo derivatization2 to form esters, acetals, hemiacetals, and esters (including halides) of such hemiacetals. Pen ta-0-ace tyl- l-chloro-l-deoxy- 1- 0- methyl - aldehydn-~-galactosealdehydro13 (compare IV), penta0 - acetyl - 1- 0 - methyl - 1- thioethyl- aldehydo - Dgalactose aldehydrol (111) and penta-0-acetyl-lbromo-l- thioethyl- aldehydo - D - galactose aldehydrol were first described by Wolfrom and coworkers, but a hetter method of synthesis (I -+ IV) for the bromo analog (IV) is the Gauthier bromination method5 as adapted by Weygand and associates.6 Compounds 11, 111, and IV may exist in C-1 epimeric forms but so far only one of each has been encountered.
I
.
(CHOAC)~ I
CH,OAC I
Br?
CH30H
I
AezCO3
(CHOAC)~ +
II/N \
0
Q
I CUOCH3
I
CHOCH3 1
(CHOACIr
I
CHOCHj I
ICHOACL
I
ICHOHI.
I
CH20AC
?nI
I
I
NUAc
CUpOAC
pm
CHpOH
x (Two forms, epimeric on C-1 )
CH~OAC
I1
to that described by Davoll and L0wy7 for the synthesis of cyclic nucleosides. Ethanolic picric acid Hk(SC2H5) HCBr(OCH3) removed the N-acetyl group from the sirupy product VI, in a manner similar to the N-debenzoylaI Br? I (CHOAC)~ (CHOA4c)4 resulting in the tion by Parikh, lT70lff, and I I of crystalline penta-O-acetyl-l-(9-adeformation CH20Ac CH~OAC n yl picrate) - 1-deoxy- 1-0-methyl-nldeh~do-D-galacI11 IV tose aldehydrol (1711). Removal of the picrate group We wish to report herein the use of this active gave crystalline penta-O-acetyl-l-(9-adenyl)-l-debromo derivative of aZdeh~/do-D-galactoseto prepare oxy-l-O-methyl- aldehydo - D - galactose aldehydiol acyclic analogs (IX) of a nucleoside. The corre- (VIII). The sharpness of its melting point indicates sponding l-chloro derivative3was also employed but that this material is probably a single sitbstnnw. its use resulted in poorer yields. Penta-O-acetyl-1- Deacetylation of VI11 with a boiling solution of nbromo- l-deoxy-l-0-met hyl-nldehydo-D-galactose al- hutylaniine in methanol produces two products. dchydrol (IT’) was condcnpcd with 6-acetamido-9- This observation leads to the conclusion that the chloromercuripurine (V) by a procedure analogous penta-O-ncetyl-l-(9-adenyl)-l-deoxy-1 - 0 -methylUldch~do-D-galartosealdehydrol is either a mixture ( 1 ) Supported by Grant S o . C Y 32311(C3), the I k p a r t ment of Health, Cducation, and Wrlfurc, Public H t d t h of l-epimers or that a partial inversion of carbon one orciirs during thc dearctylation resulting in a Service, Xatioiid Institutrs of Hrsalth, Bctliesdit 11, X l t l . ( R . F. Project 759C). Preliminary conimunic.:rtion: /I h- pair of l-epimers. T\-e designate these compounds: stracls Papers Am. C h e m Snc , 137, 3D (1WiO). First Form, m.p. 201-203.tjo, [Cy]: f24.40 (water); ( 2 ) 1f.L. Wolfrom in “Orpnnic Clieniistri,” H. Gilinaii, and Second Forin, m.p. 131-134O, [ a ] : -23.G’ editor, John Wiley and Sons, Xew York, 2nd edition, 1!)13, (water). Thew substances are a new type of nucleoVol. 11, pp. 1575-1881. ( 3 ) hl. L. TTolfrom and n.I . TYcishlat, J . A m . Cheni. side analog. They are derivatives of an acyclic heniiOCHI
-
Soc., 62, 878 (1940). (4) RI. L. IVolfrom, D. I. IYeishlat, anti ,\. R. Hanze, ,I. Am. Chem. Soc., 62,3246 (1940). (5) C. Gauthier, ilnn. pharm. frang., 1 2 , 281 (1954). (6) F. Weygand, H. Ziemann, and H . J Bestmann, Chem. Ber., 91, 2534 (1958).
( 7 ) J . Dnvoll and B. A . Loivv, J . .im. Chem. SOC.,73, 1650 (1951). (8) J. R. Parikh, lf. E . Kolff, and A. Burger, J. -4m. Chem. Soc., 79,2778 (1957).
13.55 in, 12.22 w, 9.21 m, 7.08 s ( 2 ) ,6.77 111, 6.08 vs ( l ) ,5.61 vw, 5.34s (3), 5.06 m, 4.87 vw, 4.71 m, 4.61 vw, 4.39 m, 4.20 vw, 4.00 s, 3.87 vir, 3.73 vw, 3.62 vir, 3.51 m, 3.36 m , :3.22 V W , 3.12 \I,, 3.O(i w. : i d . C!nlcd. tor C(??H29S5Oil:CI, 48.97; H, 5.42; N , 12.98. Found: C, 48.42; H, 5.42; K, 12.71. 1-(9--1den~l)-l-deoxy-l-0-methyl-aldeh~do-~-qnlactose alrlchydrol, Jrst f o r m (1x1.Penta-O-acet~yl-~-( D-udenyl)-I-deosj~I -O-niethyl-aIdehi/do-n-galactose aldrhydrol (VIII, 1.3 g.) tws r c f l u x d in 60 ml. of absolute ethanol containing 5 ml. of /1-hiitylaniine.~5~ ls The solution was concentrated to :i sirlip I)? repeatrd evaporation, under rrduced pressure, of its :ibsolute nirthanolic solution. Thcl dry residue was extracted with chloroforin to remove unrrac.ted inatrrial, and dissolved in water, decolorized with carbon, evaporated under reduced pressure, and dried by repeated evaporation of its niethanolic solution. T h e dried solid was triturated with warm ethanol EXPERIMESTAL and filtered. The filtrate upon evaporation produced a solid; Penta-O-acetyl-l-(9-aden~/l picrate)-1-deozy-1-0-methylal- yield 530 mg. (67'A). Recrystallization from methanol produced 1-(9-adenyl)-l-deoxy-l-O-methyl-aIdehydo-~-galactosc dehydo-D-galactose aldehydrol (VII). A mixture of 12.7 g. of 6-acetamido-9-chloromercuripurine ( V ) , 9 14.0 g. of cadmium wldehydrol ( I X ) , First Forin; yield 360 mg., 1n.p. 201-202.5O, [CY]:: 24.4' (c, 0.5 watcr); absorption spectra data'3: carbonate, 5 g. of Celite,'O and 425 ml. of xylene was azeo260 mp; Ai:; 2.85, 2.95, 3.10 p (OH, S H ) , 6.00, 6.10, tropically dried by distillation of 100 ml. of the xylene. T o 6.20, 6.30, 6 . 7 8 (NH,, ~ XH, and purine ring); 7 . 3 0 p (methyl this hot suspension, 15 g. of penta-0-acetyl-1-tromo-1of methyl e t h r r ) ; 8.00, !1.:35, tieoxy-1-0-methyl-aldeh ydo-D-galactose aldehydrol (1V)B was hydrogen); 8.34 p (C-0-C added with stirring.' T h e mixture \vas refluxed 3.5 hr., filtered 9.70 p ((>--.o-C, C-OH); x-ray powder ditrraction datald: hot, and the filtrate concentrated t o a sirup. T h e residue and 7.23 s, 6.09 ni, 5.66 in, 5.1s s , 4.8:3 ni, 4.48 vs ( l ) ,4.30 w, filter cake were extracted n-ith hot chloroform. The chloro- 4.12 vw, 3.93 vw, 3.64 s ( 2 ) ,3.54 m, 3.47 m, 3.38 V F , 3.28 vs form solution was washed with 30y6 potassium iodide solu- (3), 3.16 vw, 2.93 V W , 2.84 VW, 2.65 w, 2.58 w)2.37 vw, 2.30 tion] with water, dried with anhydrous sodium sclfate, and v\v, 2.32 w. Anal. Calcd. for C , ~ H ~ , S S C, O ~43.77; : H, 5.82; S , 21.27; r reduced pressure to a glassy solid ( V I ) ; ). T h e corresponding I-chloro derivative3 OCH,,9.43. Found: C:, 43.99; H , 5.77; N. 21.69; OCH3, 10.18. ployed b u t its use resulted in much lower These data indicate the suhstancr to the higher rotating yields. A solution of 4 g. of the glassy product ( V I ) in 80 nil. of epimer of 1-(9-adenyl)-l-deoxy-l-O-methyl-akfrhydo-D-g:cethanol was boiled with 11 ml. (1.1 moles) of a 10c;i ethanol lactose aldehydrol ( I X ) which F e designate the First Forni I-( deny1)-1 -deoxy-1-0-methyl-aldehydo-D-galactose aldrsolution of picric acid8 for 1 min. and the solution was cooled. The resulting yellox precipitate was filtered and washed with hydrol, second form (IX). Penta-O-acetyl-l-(9-adenyl)-ld~oxy-l-O-methyl-a2dehydo-o-galactosealdehgdrol IVIII, 1.1 a little absolute ethanol; yield 3.77 g . (717;). This material g.) was deacetg!ated as described above in the preparation of was chromatographed, in two 0.7-g. portions, on a column" (100 x 44 mm.) of Micro-Cel C.lo Development with 150 nil. the First Form. The dry residue was redissolved in methanol, of benzene-ethanol (5: 1 by vol.), produced one main yellow decolorized with carbon, and the colorless filtrate was allowed zone. The sectioned zone was eluted with acetone acd the t o evaporate a t room temperature. The initial fraction of product obtained on solvent removal was crystallized from crystals was separated; yield 200 mg. (30%).Recrystallization ethanol by evaporation at room temperature t o produce from ethanol gave I-(9-adeny1)-l-dcoxy-l-O-methyl-nldehydo-D-galactose aldehydrol (IX), Second Form, m.p. 1:31 bright yellow crystals; yield 250 mg. of penta-O-acctyl-l-(9I N o , [ C Y ] ;-23.6" : (c, 0.6 water); absorption spectra datal3 nladenyl picrate)-l-deoxg-l-O-methyl-aldeh~/do-n-ga1iictosc~ 260 mp; ;:A: 2.92, 3.10 (OH, N H ) ; 6.00, 8.10, 6.35, dehydrol (VII), dcc. 287-280'. 6 . 7 5 (~S H , , S H , and purine ring); 7.25 i.( (methyl hydrogen I ; .4nal. Calcd. for C28H82x8018: N, 14.57. Found: N, 15.61. 8.30 p (C-0-C); 9.10, 9.40, 9.70, 9.82 p (C--0-C, (:--Penta-0-acetyl-1-( 9-adenyl )-I -deox y-1-0-meth yl-nldeh ydo-1)OH); x-ray pon-der diffraction data'l: 12.95 vw, 7.90 m, 7.25 galactose aldehydrol (VIII). A solution of 2.4 g. of penta-0acetyl- 1-(9-adenyl picrate)- I-deoxy- 1-&met h yl-aldeh ydo-n- s, 6.99 m, 6.43 m, 6.04 m, 5.75 vw, 5.58 w, 5.28 w, 4.98 ni. galactose aldehydrol (171)in 500 ml. of 5070 aqueous ace- 4 . 6 6 w v , 4 . 4 0 w , 4 . 0 9 m3, . 9 2 v s ( l ) , 3 . 8 0 s , 3 . 6 7 w , 3 . 5 : ~ v s 1 2 ) , tone was passed through a 30 X 420 mm. column of Do~ves-1 3.36 vs ( 3 ) , 3.24 vw. Anal. Calcd. for Cl?Hl,Ns06: C , 43.77; H, 5.82; K , 21.27. (carbonate form) anion exchange resin12 and the column was OCH3, 9.43. Found: C, 43.55; H, 5.93; K,21.09; OCH:, 9.10, washed with 700 ml. of thc solvent. T h e solution \vas evaporated to 350 ml. and extracted thrice with 75 ml. of rhloroform, T h e extract was dried with anhydrous sodium sulfate ( 1 3 ) The ultrsviolct absorption analyses were made on and concentrated t o a sirup. T h r material crystallized from i t Cary Rerording Spectrophotoinptm-, Alodel 10, Applird absolute ethanol; y k l d 1.11 g. Two rcw),stallizations from Physics ('orp., Pasadena, Calif. The infrared spectral datii !)5'7; ethanol produced fine necdrs, m.p. 191-1F)2°0, [ C Y ] '," were obtained with an Infrared Recording Spectropho-25" (c, 0.8 chloroform); absorption spectra datal3: tometer] Model B, Baird Associates, Inc., Cambridge, Mass. 260 mp, 3.00, 3.15 p (NH2, NH), 5 70 ,u (ester carhonyl), Structural assignments were made following TI'. B. S e e l y , 5.98, 6.10, 6.22, 6.80 p ( N H 2 , NH, and purine ring); 7.30 p Advances i n Carbohydrate Chem., 12, 13 (19,57) and B. R . (methyl hydrogen); 8.22-8.38 p (broad C-0-C of ace- Baker and associates.8 x-ray powder diffraction data": tates), 9.70 p (C--0-C); (14) Interplanar spacing, A, C u K a radiation. Relative intensity, estimated visually: s, strong; m, medium; w , weak; v, very; three strongest lines numhered in order of inweas(9) B. R . Baker, Kathleen Hewson, H . Jeanette Thomas, ing strength. and J . A. Johnson, Jr., J . Orq. Chem., 2 2 , 954 (1957). (15) L. Goldman, J. W. Marsipo, and R . B. Angicr, J . (10) A product of Johns-Xlanville Co., Xew York, N. Y . Am. Chem. Soc., 78,4173 (1956). (11) W. H. RlcXeely, W.W. Binkley, and M. L. \T'olfrom, (16) E. J . Rcist and B.R. Bnkcr, J . Org. Chem., 23, 1083 J . A m . Chem. SOC., 67, 527 (1945). (12) A product of the Dom Chemicd Co., LIidland, Mich. (1958).
acetal while the nucleosides are derivatives of cyclic hemiacetals. We also wish to report the syiithebis and characterization of 9-P-u-galactopyl.anosyladeninc. which was obtained by the condensation of tetra-o-acetyla-D-galactopyranosyl bromide with 6-acetainido-9chloromercuripurine (V) . The product was purified through g-(tetra-O-acetyl-P - 11- galactopyraiiosy1)adenine and its picrate, both crystalline substances. Iternoval of the picrate and acetyl groups resulted in crystalline 9-(P-D-galactopyranosyl)adenine, characterized further by its crystalline picrate.
+
'I'htw ilat:i irdir:itcs that this substsiicc is t,h(,lower rotating 1-epimw of I-(9-atlenyI)-I-deosy-l-O-niethyl-aIdehydoii-y:il:ictosc alclc~hydrol ( I S ) , hwcin clcsignated as thc Swond Form. !I-( ? ' e t , . t c - O - u c , c l ! / l - P - U - Y ( I ( U L . l o / J I / I . picrule. .i mixture of 28.0 g. of 6-acctaniido-'3-clIloromrrcurip1irine (V),@ 28.0 g. of cadmium carbonate, 5 g. of Celite,IO and 425 ml. of xyltAnc wns dried azeotropically by distillation of 100 ml. of thv xylcric~.To this miyture \ v : i ~atltletl, : i t 90" :inti wit>hstirriiig, :L solrition of 28.0 g. of tetr:i-(i-:ic.r,tyl-n-i,-galnc.topyr:itiosj.1 trroiiiitl(~~7 i i i :jO i n l . of syltno. 'I'hr, mist iirr \vas refliised for 3 hr., cwcnti:dIy according t o thc. procrdrire of 1)avoll and I,o\vy,i filtcrtd hot, and tho filtr:itcl \vas conccntratrd rinder wdiicrd prwsrirc. TIN.filter calcc~:ind resitliie i v w e extracted \vith hot chloroforin, the chloroform solution \vas washed \vith 30';; acjnt'ous potassium iodide, watrr, dried Kith anhydrous sodium sulfate, and concentrated under reduced pressure to a sirup; yield 21.0 g. of crude G-acetamido-8(tetra-0-acet~l-p-u-galactopyranosyl)piirine. The crude product (13.1 g.) \vas convwtcd to the picrate by boiling for I min. v i t h 87 ml. of lOyo picric acid in cethano1.s The picrate of 9-(totra-0-acf,tyl-P-D-galactopyrano crystallized upon cooling; yicLltl 12.4 g. \vas f'urthcxr piirificd by coliinili chromxt 0.8-g. portions of matwial 011 (100 x 44 m m . ) columns of .\Iicro-('csl (:lo iisiiig 150 inl. of Iirnzcne-rth:inol ( 5 :1 by vol.) :IS dc~vc~loper. Tht. matrrial in the bright yellow zone. \vas (,luted with actltorie and crystallized from ethanol as (Irxrj) j.clloiv riec:tllcs, ticc. 281-287". .trial. Calcd. for C ( ? ; H ? g S & 1 6 : S , Iti.14. Found: S , 15.90. D-( T e l r a - O - a c e t ~ l - P - D - g a ~ a c ~ o p ~ r a n o s ? / ~A ) umixture ~e~~z~e. of !~-(totr:~-0-:~cetyl-~-I,-galnctopyranos~l):~tlenine picrate ( i . lg.) i v w convrxrtd to the nucleosidr by trr:itmrnt with J)o\vcx-l as drscrihrtl a h o w for the acyclic derivative. Th(. rwrilting sirup ( 3 . 7 g., 75'~,)\v:w concentrated under reduced prwsiirc t\vice from ethxnol, tlissolvrd in 25 ml. of ethanol, :incl :illo\vtd to evxporatc from an o p m t m k e r :it room temi1lix:ttion occiirrcd in 24 lir. The material was filtertrd and xashcd with cold methanol; yield 1.13 g. ( 2G(!A) of O-( t rt r:i-O-acct yl-b-n-ga lactopyranos?;l)adenin~ .-___ -.
( I T ) H . Ohlr, \T..
llarerelq and
IT;.
Bourjau, Ber., 62,
8x3 ( 1929).
ItecrystalliziLtion from methanol produced fine white 11(vd e s ; m.p. 212-213.5", [a]1,4$7.3" (c, 2.6 chloroform); absorption spectra datal3: X ~ ; ~ s o R2-48 mp, 2.88,3.05p ( N H , S H ? ) , 5.G5 p (C-0 of acetates), 5.95, 6.15, 6.70, 6.!)6 p ( S H , aiid purine ring), i . 2 8 (niethyl ~ hydrogens), 8.05-6.22, 9.15, 9.45 p ( C 4 - C ) ; x-ray powder diffraction datal4: 11.07 vs ( l ) ,9.72 vw. 8.68 vw, i.86 m, 6.76 in, 6.38 R, 6.04 w , ,5.65 ni, 5.10 s (Z),4.82 m ( 3 ) , 4.31 in,4.19 m, 3.94 w, 3.85 vw. .Anal. Calcd. for C I ~ H ~ S C, ~ O 49.02; ~ : H, 4.98: X, 15.15, I~'ourit1:C, 48.05; H, 4.40; S , 15.52. D-P-u-Grr/actupUrarto.s~/~[~e~~?te. .A solution of 24.0 g. 01 crude 6-acr~talnido-!l-(tetra-0-acetyl-p-D-galactopyranos).l)purinc in 225 ml. of dry niethanol arid 35 nil. of n-butylaminv \vas refluxed for 5 hr. T h e solution \vas concentrated to 50 ml. under reduced pressurc and held :it 0 " until precipitation occurred. The precipitate \vas filtered and washed with cold methanol, chloroform, and ethrr; yield 9.6 g. (Gi.5S).The solid was dissolved i n water, decolorized with carbon and coriccJntratrd to a sirup whirh crystallized. T h e mass rvas triturated with methanol, filtered, and dried; m.p. 192-195". Further purification was c~ffectedby treatment with hot 105; ethanolic picric acid to form a rrystallinc F)-(P-D-galactopj.ranosy1)adeninr picrate; m . p . 230-232'. .lnal. Calcd. for C17H18S&1?: C, 38.79; H, :i.4,5: S , 21.2:J. Found: C, 38.83: H , 3 . 6 5 ; S , 19.98. Regenrration of the nucleoside from the picrate with I )owcs-l (earbonatc iorin)l2 anion exchange resin by the method tlescribed above for t h e acyclic picrate derivative gave fine colorless iieedles, m.p. 198-200", [a]'; +95.5' ( c , 0.5 water); absorption spectra datal? A,",: 260 m p ;: :A: 2.00, 2.98 p (OH, NH), 8.02, ti.24, 6.32, 6.78 p (XH?, K H , and C-OH); x-ray purine ring), 9.20-9.50, 10.0 p (C-0-C, powder diffraction datn": 7.65 w, 7.00 vw, 6.59 ni, 6.35 m, .5.88 w, 5.20 vw, 1.88 vs ( l ) ,4.60 \v, 4.32 vw, 4.18 w, 4.08 vw, :%.!I7 w, 3.60 ni, 3.50 m ( 3 ) , 3.42 K, 3.36 TV, 3.26 s, ( 2 ) , 3.17 \v, 3.07 VW, 2.!14 w,2.89 w. A n a l . Calcd. for Cll Hl5rsOj: C, 44.43; H, 5.08; S , 23.51. Found: C , 43.85; H , 5.36; S , 23.31. Identicnl material was also obtained on deacetylatioii, with boiling methanolic n-butylamine, of F)-(tetra-O-acet3-1-8D-galactopyranos~l)adpninr hy the same procediirr iisrrl t o deacetylate VIII. Cor,r;>ims 10, OHIO
[ ~ ~ ( I S T R I R I ~ T IFROM OS THE D E P A R T M E N T O F AkCRICULTURAL BIOCHEMISTRY, H A W A I I .&ORICUI.TUR.AL E X P E R I M E N T sT.ITIO?J
UNIVERSITY OF HAWAII]
The Constitution of a Galactomannan from the Seed of Leucaena glaiical -4.51. UNRAU2 Received January 8, 1961 A galactoinannan, composed of 57% mannose and 43% galactose, \vas isolated in Z5yo yield from the seed of Leztcaena qlauca. The po1ysacch:iride had a n average D.P. of 150 and gave highly viscous aqueous solutions a t low solute concentrations. Pcjriod:itc t1cgr:idation revealed t h a t some mannose residues and a smaller nuniher of galactose residues were not attacked. Hydrolysis of the methylated gum showed t h a t galactose occupied mainly terminal, nonreducing positions and the found indicated the presenrr of some major branch-points. considerable qusntity of 2,:3,4,6-tetra-O-methyl-~-mannose Flirther evidence t h a t some 1,3-linkages were present was ohtained. DISCUSSIOS
The W d of Le"caena gzauca (better known as koa honk in the Hawaiian Islands) has an interesting -.
__
chemical composition. Among the nitrogenous constituents found in the seed, a-poisonous-amino acid kllown as mimosine3 1s present in relatively large quantities. The isolation of this amino acid whirh
( 1 ) Published with approval of the Director, Hawaii
:ie;ricultriral Experiment Station, as Technical paper no. 508. ( 2 ) Prtsent addrms of author: Department of Chemistry, 17riiversit\,of UritishColririitiitt, Vancoriver 8, Canada.
( 3 ) J. Renz, 2. Physiol. (-'hem., 244, 153 (1936); H . Nienburg nnd K. Taubock. %. Physiol. Chem., 250, 80 (1937); 1) k'ostermans, R e v . T r w . Chim., 6 5 , 319 (1946).
